The Effect of γ-secretase Inhibitor Combined with BMSC on the aGVHD in Mice Model / 中国实验血液学杂志

OBJECTIVE@#To establish the aGVHD mouse model，and investigate the regulatory effect and its mechanism of low-dose GSI combined with BMSC on aGVHD mice.@*METHODS@#C57BL/6 (H-2b) and BALB/c (H-2d) were selected as donor and recipient of allogeneic transplantation to establish the aGVHD mouse model. BALB/c mice were randomly divided into 6 groups, which were the bone marrow cell infusion after irradiation (BM) group; the bone marrow cells + spleen cells after irradiation (BM+SC) group; the bone marrow cells + spleen cells + DMSO (BM+SC+DMSO) (transplant control) group; bone marrow cells + splenocytes +GSI after irradiation (BM+SC+GSI) group; bone marrow cells + spleen cells + bone marrow mesenchymal stromal infusion after irradiation cell (BM+SC+BMSC) group; bone marrow cells + spleen cells + bone marrow mesenchymal stromal cells +GSI infused after irradiation (BM+SC+BMSC+GSI) group. The mice in the two groups containing GSI were intraperitoneally injected with GSI at 5 μmol/kg on day 1, 2, and 3 after transplantation with DMSO as a control. The general conditions, survival time and hematopoietic recovery of mice were observed, cytokines were detected by ELISA, and histopathological changes were detected by immunohistochemistry. The effects of low-dose GSI combined with BMSC on hematopoietic reconstruction and aGVHD development after allo-BMT were investigated.@*RESULTS@#The survival rate of the mice in BM+SC+BMSC+GSI combination group was 80% during the observation period, which was significantly higher than that in the other groups; the incidence of aGVHD was reduced in the BMSC GSI or their combination groups after 21 days of transplantation. GSI could partly promote the recovery of leukocytes, and show no significant delayed effect on the recovery platelets. Moreover, the level of Th1 cytokines (IFN-γ) in BM+SC+BMSC+GSI combined group was lower than that in BM+SC+GSI group (P<0.01), the level of Th2 cytokines (IL-4) in the combination group was higher than that in BM+SC+GSI group (P<0.01), also the level of IL-17 was significantly lower than that in the corresponding control group (P<0.001).@*CONCLUSION@#Low dose GSI combined with BMSC can promote hematopoietic reconstruction and regulate cytokines secretion including IFN-γ, IL-4 and IL-17. GSI combined with BMSC achieve the goal of synergistically inhibiting the occurrence and progression of aGVHD.

Inhibition of invasion and metastasis of human tonguesquamous cell carcinoma Tca8113 cells by Src kinase inhibitor PP2 / 西安交通大学学报(医学版)

Objective: To investigate the inhibitory effects of Src kinase inhibitor PP2 on migration and invasion of Tca8113 cells. Methods: Tca8113 cells were cultured for 24 h with 5 mol/L, 10 mol/L and 20 micron mol/L of Src kinase inhibitor PP2. The effects of PP2 on the invasion and migration of Tca8113 cells were measured with Transwell chamber and scratch method, respectively. Results: After the treatment with PP2 for 24 h, the expression of p-Src in 5, 10, 20 μmol/L of Src kinase inhibitor PP2 treatment groups was significantly lower than that of the non-drug treatment group (all P<0.05). The number of Tca8113 cells in the non-drug treatment group and the 5, 10, and 20 μmol/L of Src kinase inhibitor PP2 treatment groups was (232.76±28.65), (186.53±21.34), (129.18±17.96), and (37.82±12.41), respectively; the number of migratory cells was (259.38±25.27), (193.45±20.18), (143.24±18.04), and (32.94±14.39), respectively, the cell migration rate was (11.51±0.84)%, (8.06±0.51)%, (5.13±0.57)%, and (3.18±0.12)%, respectively; the overall difference was statistically significant (F=73.852, 85.687, 48.157, all P=0.000). It had a negative correlation with PP2 dose. Conclusion: Src kinase inhibitor PP2 can effectively inhibit the invasion and migration of Tca8113 cells in the concentration-dependent manner, and it may have certain clinical value in the treatment of human tongue squamous cell carcinoma.

Research Progress on Notch Signal Pathway in Acute Graft-Versus-Host Disease -Review / 中国实验血液学杂志

The Notch signaling pathway is a highly conserved cell signaling system that plays an essential role in many biological processes. Notch signaling regulates multiple aspects of hematopoiesis, especially during T cell develop-ment. Recent data suggest that Notch also regulates mature T cell differentiation and function. The latest data show that Notch also plays an essential role in alloreactive T cells mediating acute graft-versus-host disease (aGVHD), the most severe complication of allogeneic hematopoietic stem cell transplantation (allo-HSCT). Notch inhibition in donor-derived T cells or blockade of individual Notch ligands and receptors after transplantation can reduce GVHD severity and mortality in mouse models of allo-HSCT, without causing global immunosuppression. These findings indicate Notch in T cells as an attractive therapeutic target to control aGVHD. In this article, the pathophysiology of aGVHD, the Notch signal pathway and aGVHD are reviewed.

Detection of microvesicle miRNA expression in ALL subtypes and analysis of their functional roles / 华中科技大学学报（医学）（英德文版）

Microvesicles (MVs) are the heterogeneous mixtures of vesicles. MVs released by leukemia cells constitute an important part of the leukemia microenvironment. MVs might act as important reservoirs of microRNAs (miRNAs). It is worth evaluating whether MVs possess some unique miRNA contents that are valuable in understanding the pathogenesis. In this study, we investigated the miRNA expression patterns of Nalm-6-derived MVs, Jurkat-derived MVs and normal cell-derived MVs using miRNA microarrays. The potential target genes regulated by differentially expressed miRNAs were also predicted and analyzed. Results demonstrated that 182 miRNAs and 166 miRNAs were differentially expressed in Nalm-6-MVs and Jurkat-MVs, respectively. Many oncogenes, tumor suppressors and signal pathway genes were targeted by these aberrantly expressed miRNAs, which might contribute to the development of B-ALL or T-ALL. Our findings expanded the potential diagnostic markers of ALL and provided useful information for ALL pathogenesis.

Detection of microvesicle miRNA expression in ALL subtypes and analysis of their functional roles / 华中科技大学学报（医学）（英德文版）

Microvesicles (MVs) are the heterogeneous mixtures of vesicles. MVs released by leukemia cells constitute an important part of the leukemia microenvironment. MVs might act as important reservoirs of microRNAs (miRNAs). It is worth evaluating whether MVs possess some unique miRNA contents that are valuable in understanding the pathogenesis. In this study, we investigated the miRNA expression patterns of Nalm-6-derived MVs, Jurkat-derived MVs and normal cell-derived MVs using miRNA microarrays. The potential target genes regulated by differentially expressed miRNAs were also predicted and analyzed. Results demonstrated that 182 miRNAs and 166 miRNAs were differentially expressed in Nalm-6-MVs and Jurkat-MVs, respectively. Many oncogenes, tumor suppressors and signal pathway genes were targeted by these aberrantly expressed miRNAs, which might contribute to the development of B-ALL or T-ALL. Our findings expanded the potential diagnostic markers of ALL and provided useful information for ALL pathogenesis.

Inhibitory effects of gamma secretase inhibitor on human multiple myeloma xenograft mouse model / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the tumor growth inhibition of gamma secretase inhibitor MRK003 on human multiple myeloma xenograft mice by inhibition of AKT and Notch1 expression.</p><p><b>METHODS</b>NOD/SCID mice were injected with human multiple myeloma cell lines RPMI8226 to establish a xenograft mouse model. Mice were randomized into two groups：the experimental group were injected with MRK003 at a dose of 5 mg× kg⁻¹×d⁻¹ for 14 days; the inhibitor was replaced by an equal saline in the control group. Mice were sacrificed by cervical dislocation on the next day after the last injection and tumor tissue was removed to detect the expression of Notch1 and AKT by immunohistochemistry.</p><p><b>RESULTS</b>After subcutaneous injection with RPMI8226, mice had tumor formation in 5-7 days and the largest tumor block in 10-12 days. Before RPMI8226 injection, the mean sizes of tumor block in the experimental and the control groups were 509.2 mm³, 511.2 mm³(P>0.05). 9 days after injection, the mean sizes of tumor tissue in the experimental and the control groups were 636.6 mm³, 691.2 mm³(P<0.01). On the next day after the last injection, the tumor sizes of the experimental and the control groups were 683.5 mm³ and 1798.7 mm³(P<0.01). The size of tumor block in the experimental group was significantly smaller than that of the control group(P<0.01). Immunohistochemistry staining showed that the positive expression rates of Notch1(11.1%, P<0.01) and AKT(13.3%, P<0.01) in experimental group were significantly decreased compared with the control group(Notch1: 95.6%; AKT: 93.3%). Western blot results showed that Notch1 and AKT protein in experimental group were significantly lower than those in the control group.</p><p><b>CONCLUSION</b>MRK003 could inhibit the tumor growth of human multiple myeloma xenograft mice by downregulated expression of Notch1 signaling pathway.</p>

Study of fibrinolysis inhibitors in 117 acute leukemia patients / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the clinical significance of fibrinolysis inhibitors including thrombin activatable fibrinolysis inhibitor(TAFI), plasminogen activator inhibitor (PAI) and alpha2-plasmin inhibitor (alpha2-PI) in acute leukemia (AL).</p><p><b>METHODS</b>PAI-1 antigen and TAFI antigen were investigated by enzyme-linked immunosorbent assay and PAI activity, alpha2-PI activity, TAFI activity by chromatography substrate assay in 117 AL patients and 50 normal controls.</p><p><b>RESULTS</b>1) alpha2-PI activities in AL patients were reduced, especially in ALL patients [(96.8 +/- 21.2)%]; 2) PAI-1 antigens in AML patients [(37.8 +/- 9.2) microg/L] were significantly higher than that in normal controls [(33.8 +/- 4.9) microg/L]; 3) PAI-1 antigens in APL [(37.8 +/- 9.0) microg/L] and AML-M5 patients [(39.9 +/- 11.6) microg/L] were higher and TAFI activities in APL patients [(13.3 +/- 4.8) mg/L] were lower than that in normal controls [(16.9 +/- 2.6) mg/L]; 4) PAI-1 antigens of relapsed/refractory patients (39.6 +/- 11.6) microg/L) were significantly elevated; 5) TAFI activities in bleeding patients [(13.2 +/- 5.3) mg/L] were significantly lower than that in normal control as well as non-bleeding patients (17.0 +/- 4.6) mg/L); 6) The severity of bleeding was negatively correlated with TAFI activity (r = - 0.276, P <0.05).</p><p><b>CONCLUSIONS</b>Hyperfibrinolysis caused partially by decrease of alpha2-PI and TAFI activity takes part in the pathogenesis of bleeding in AL.</p>

Notch signaling in human breast cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the role of Notch signaling in human breast cancers, the expression of Notch1 and its ligand JAG1 in human breast cancers and their relationships with clinical stages of breast cancers were analyzed.</p><p><b>METHODS</b>RT-PCR was used to detect the expression of Notch1 and JAG1 in 62 breast cancer specimens and 22 normal breast tissues at the margin of tumor sections, and the statistical difference of expression rates and standardized coefficient between the two groups were analyzed. To compare the expression intensity of Notch1 and JAG1 at different development stages of the illness and at different stages with or without axillary node metastasis.</p><p><b>RESULTS</b>The expression rate and standardized coefficient of Notch1 in human breast cancers were significantly higher than those of normal breast tissues at the margin of tumor sections. The expression rate of JAG1 in human breast cancers was 15%, while JAG1 was not detected in normal breast tissues at the margin of tumor sections. The standardized coefficient of Notch1 in cases with axillary node metastasis was significantly higher than that in cases without axillary node metastasis. The standardized coefficient of Notch1 at stage I was significantly lower than that at stage II, and stage II was significantly higher than stage III. There was no statistically significant difference between stage I and stage III.</p><p><b>CONCLUSION</b>Notch1 and JAG1 are highly expressed in human breast cancers, indicating that the aberrant expression and activation of Notch1 may be related with tumorigenesis of human breast cancer. Notch1 may play different roles at different developmentl stages of human breast cancer.</p>

Antagonistic Effects of Organic Extracts of Fruit of Chinese Wolfberry on Chromium (Ⅵ)-induced Mutation in Human Peripheral Blood Lymphocytes / 环境与健康杂志

Objective To study the antagonistic effect of organic extracts of Fruit of Chinese Wolfberry (FCW) on chromium (Ⅵ) _induced mutation. Methods Using unscheduled DNA synthesis (UDS) assay, the effects of 0.02 g/ml, 0.01 g/ml and 0.005 g/ml organic extracts of FCW on UDS induced by potassium bichromate [0.31 ?g/ml Chromuim (Ⅵ)] in human peripheral blood lymphocytes were determined. The effects of the order of adding organic extract of FCW and potassium bichromate into cultured lymphocyte suspension on UDS were observed at the same time. Results The value of CPM revealed a significantly higher level in positive control group exposed to 0.31 ?g/ml chromium (Ⅵ) compared with that in negative control group exposed to disstilled water (P